Demonstration of mass transfer and pH effects in a nitrifying biofilm

A bench-scale nitrifying trickling filter (surface AREA = 0.5 m²) was developed to permit evaluation of diffusion of oxygen within a biofilm, the pH dependence of ammonium oxidation and external mass transfer. In addition, a biofilm model was developed and verified for homogeneous nitrifying biofilms of varied thickness and for thin nitrifying biofilms covered by heterotrophic biofilms. The model uses literature values for the pH dependence of Monod coefficients for Nitrosomonas and Nitrobacter.

The diffusion coefficient of oxygen in the biofilm was found to be 40–80% of the value in pure water. Due to mass transfer resistance, the biomass ·sees” a lower pH than is measured in the water film passing over it. The surface uptake rate of ammonia is used as an indicator of pH gradients within the biofilm system. With the help of oxygen limitation experiments, the location of nitrifying biomass within mixed biofilms (heterotrophic, autotrophic) can be determined.

The biofilm model predicts ammonium uptake rate of a trickling filter as a function of the bicarbonate concentration in the water film.     

Aerobic treatment of piggery waste

The operating characteristics of laboratory waste treatment systems were studied during the aerobic degradation of pig excrement at different loading rates and temperatures. The treatment systems were of two types: one was operated with floc formation and gravity separation of liquid and suspended solid effluents; and a second was operated without floc formation or separation of the effluent into liquid and solid fractions.

With an operating temperature of 15°C the parameters most affected by loading rate were (1) the concentrations of suspended solids and chemical oxygen demand in the liquid effluent; (2) the pH value of the mixed liquor; (3) nitrification; (4) the BOD of the supernatant from the mixed liquor; and (5) output of suspended solids as a percentage of input.

The concentrations of suspended solids and chemical oxygen demand in the liquid effluents were little affected by loading rates in the range 0·05-0·15 g SS g MLSS⁻¹ d⁻¹ (0·02-0·06 g BOD g MLSS⁻¹ d⁻¹) but increased with increasing loading rate in the range 0·15-0·30 (0·06-0·12 BOD). At loading rates below about 0·17 g SS g MLSS⁻¹ d⁻¹ (0·07 g BOD g MLSS⁻¹ d⁻¹) the mixed liquors were acidic, with pH values down to 5·2, whereas at loading rates above about 0·80 (0·32 BOD) they were alkaline, with pH values up to 8·9. At intermediate loading rates the mixed liquor pH value was more variable though in general the higher the loading rate the higher also the pH value of the mixed liquor. Acidic conditions in the mixed liquors were attributed to the occurrence of nitrification, while in the absence of nitrification the mixed liquors remained alkaline. The concentration of BOD₅ in the supernatant from the mixed liquors increased with increasing loading rate from about 35 mg 1⁻¹ at a loading rate of 0·17 g SS g MLSS⁻¹ d⁻¹ (0·07 g BOD g MLSS⁻¹ d⁻¹) to about 250 mg 1⁻¹ at a loading rate of 1·30 (0·52 BOD). The output of suspended solids from the treatment systems represented about 70 per cent of input suspended solids at loading rates of about 0·15 g SS g MLSS⁻¹ d⁻¹ (0·06 g BOD g MLSS⁻¹ d⁻¹) and increased to about 100 per cent at loading rates of 0·80 (0·32 BOD). Output of chemical oxygen demand was about 60 per cent of input at the lower loading rates and 80–90 per cent at the higher ones.

Operation of treatment units at temperatures of 5 and 10°C instead of 15°C had little effect on the efficiency of degradation at loading rates in the range 0·085-0·20 g SS g MLSS⁻¹ d⁻¹ (0·034-0·08 g BOD g MLSS⁻¹ d⁻¹), but nitrification was prevented at 5°C. At loading rates of 0·77 (0·31 BOD) and 1·46 (0·58 BOD) operation at 25°C appeared to increase the amount of degradation as compared with that achieved at 15°C.

The practical implications of the results and possible future approaches to the aerobic treatment of farm wastes are discussed.     

Phosphorous limitation in an arctic river biofilm—A whole ecosystem experiment

A fourth-order arctic river was experimentally enriched with phosphate (7.7 ± 7.0 μ g 1⁻¹) to determine the effect of such a loading (equivalent to a community of 10,000 people) upon the trophically important biofilm. The effect upon a light-grown biofilm (an autotrophic/heterotrophic assemblage) and a dark-grown biofilm (predominantly heterotrophic assemblage) was determined after 28 days of colonization. Seven attributes of the biofilms were monitored, 2 autotrophic indices, chlorophyll , [¹⁴C]HCO₃ incorporation into lipids and 5 heterotrophic indices; [¹⁴C]acetate incorporation into lipids, metabolic heat output, turn-over times of microbially labile glucose and glutamate and mineralization of microbially recalcitrant ring-labelled [¹⁴C]hydroxybenzoic acid. The findings showed that the addition of phosphorus resulted in a substantial stimulation of both autotrophic and heterotrophic processes suggesting that arctic rivers of this type would be liable to cultural eutrophication.     

Denitrification with natural gas and various new growth media

Biological denitrification was investigated in an attached growth reactor system using several growth media, denitrifying cultures and natural gas (95% methane) as a carbon source. In order to establish a baseline of operation, initial experiments were conducted with a bed of 2–3 mm sand and methanol as a carbon source using a methylotrophic denitrifying culture and then the system was compared with natural gas using various methane utilising cultures. Compared to methanol, performance with methane was considerably lower. In order to improve denitrification with methane a plastic medium (Etapak, surface area 200 m²/m³) was placed above the sand bed (which increased the surface area for bacteria growth in the upper part of the bed), and a new methanotrophic mixed culture (NCIMB-code 11085) was introduced to the system. This combination resulted in a 27% higher denitrification efficiency. Experiments were continued by systematically varying the operating conditions to obtain highest denitrification using methane gas and replacing the Etapak media with different plastic media of higher surface area, but keeping the NCIMB culture unchanged. Other media tested were Pall-rings (surface area 319 m²/m³), IP-spacers (surface area 500 m²/m³) and granular activated carbon (GAC-code: Norit PK 1–3). Best results were obtained with IP-spacers which, surprisingly, are designed for use in the concrete industry rather than as a bacterial support medium. These produced nitrate removal efficiencies of up to 93% at 0.6 m/h or 55% at 1.6 m/h water filtration rates. Run times of 10 days or more to a limiting headloss of about 1.0 m,were usually achieved before “bumping” or back-washing to reduce headloss. Effluent turbidities were generally below 1.0 NTU. Tests for bacteria present with GAC media and COD removal with IP spacers were also carried out. Results are discussed with operational conditions and denitrification efficiencies achieved.     

Influence of temperature and substrate shocks on survival and succinic dehydrogenase activity of heterotrophic freshwater bacteria

The influence of temperature shocks on aquatic bacteria was studied in laboratory experiments with regard to population shifts in lake water communities of aerobic heterotrophic bacteria including coliforms. as well as in synthetic mixed cultures of psychrotolerant aquatic isolates. In addition, succinic dehydrogenase activity was used as a parameter of bioactivity. Each experiment was designed to study concomitant effects of elevated levels of organic substrates.

Simultaneous additions of complex nutrients such as 100–300 mg 1⁻¹ of meat peptone to lake water caused a synergistic action of substrate and temperature on population dynamics by depressing population densities at temperatures below 20°C. In contrast, stimulation was observed at higher temperatures particularly for the coliform group. In mixed cultures consisting of Cytophaga, Chromobacterium and Arthrobacter as important members of the heterotrophic microflora of the investigated lake water, substrate accelerated death was induced by temperature shocks beyond the maximum growth temperatures. The succinic dehydrogenase activity of a psychrotolerant strain of Cytophaga failed to show more sensitive responses to temperature shocks than did viable counts.     

Pre-acidification in anaerobic sludge bed process treating brewery wastewater

The effect of pre-acidification on anaerobic granule bed processes treating brewery wastewater was the focus of a comparison study employing two configurations, (a) a single stage upflow anaerobic sludge bed (UASB) and (b) an upflow acidification reactor in series with a methanogenic UASB. The pre-acidification reactor achieved 20±4% SCOD removal and 0.08±0.003 L of methane produced per gram of SCOD removal at a hydraulic retention time (HRT) of 0.75–4 h. Butyric acid was not detected and short chain fatty acids (SCFAs) were mainly acetic and propionic acids. The acidification ratio was about 0.42±0.02 g SCFAs as COD/g of influent COD.

Both systems’ critical loading rate to achieve 80% COD removal was established at 34–39 kg COD/m³ of total sludge bed volume per day. SCOD removal efficiency of 90±3% was achieved by both systems at an organic loading rate of 25±1 kg COD/m³ of total sludge bed volume per day, indicating that the installation of an acidification reactor had no effect in terms of the maximum granular activity, biomass granulation and the settleability of granules. At an organic loading rate of 67 kg COD/m³ of total sludge bed volume per day at an HRT of 1 h, the series system outperformed the single UASB by a removal of 62 compared to 57%.     

Kinetics of alachlor transformation and identification of metabolites under anaerobic conditions

Alachlor is one of the two most commonly used herbicides in the United States. In the environment, little mineralization of this compound has been found to occur, and metabolites of alachlor may be formed and could accumulate. The objectives of this study were to determine the rate of alachlor biotransformation and to identify the transformation intermediates formed under aqueous denitrifying, methanogenic, and sulfate-reducing conditions. Second-order biotrasnformation coefficients for alachlor were determined to be 7.6 × 10⁻⁵ (±4.0 × 10⁻⁵), 2.9 × 10⁻³ (±1.6 × 10⁻³), and 1.5 × 10⁻² (±1.4 × 10⁻²) 1 mg VSS⁻¹ day⁻¹ under denitrifying, methanogenic, and sulfate-reducing conditions, respectively. Acetyl alachlor and diethyl aniline were positively identified as transformation products of alachlor under all conditions. In denitrifying reactors aniline was identified as a product of alachlor. When acetyl alachlor was fed as the parent compound, aniline was also identified as a transformation product under methanogenic conditions. This research showed that although alachlor is degraded under denitrifying, methanogenic, and sulfate-reducing conditions, significant concentrations of several metabolites are formed and are only slowly degraded.     

Zero-valent iron pretreatment for enhancing the biodegradability of RDX

Hexahydro-1,3,5-trinitro-1,3,5-triazine (C₃H₆N₃(NO₂)₃, royal demolition explosive or RDX) is a common nitramine explosive and one of the major constituents in wastewaters from ammunitions plants. The objective of this study is to investigate zero-valent iron (Fe(0)) pretreatment for enhancing the biodegradability of recalcitrant RDX. It was hypothesized that iron pretreatment can reductively transform RDX to products that are more amenable to biological treatment processes such as activated sludge. Results of batch and column experiments showed rapid and complete removal of RDX by Fe(0) regardless of the buffering capacity. Formaldehyde (HCHO), a major reduction product of RDX, was readily biodegraded by a mixed culture. Respirometric data indicate that iron-treated RDX solution exerted substantially higher biochemical oxygen demand (BOD) than untreated RDX solution. We propose that an integrated iron reduction—activated sludge process may be a feasible option for treating RDX-laden wastewater.     

Effects of elevated operating temperatures on methanol removal kinetics from synthetic kraft pulp mill condensate using a membrane bioreactor

The feasibility of biologically removing methanol from kraft pulp mill evaporator condensate was investigated, using a high temperature membrane bioreactor (MBR). Over the range of temperatures investigated (55–70°C), a mixed culture of methanol-utilizing microorganisms could be successfully developed, using synthetic condensate as a feedstock. A maximum specific methanol utilization coefficient of approximately 0.81 day⁻¹ occurred at an operating temperature of 60°C. Over 99% of the methanol was removed from the condensate at operating temperatures of 55 and 60°C. Above 60°C, the specific methanol utilization coefficient declined sharply, indicating that at high operating temperatures, the inactivating effect of temperature on the mixed culture of microorganisms must be considered. A relatively simple model was proposed and used to estimate the effect of high temperatures on methanol removal kinetics in an MBR over the range of temperatures investigated. The operating temperature also had a significant effect on the observed growth yield. At increasing operating temperatures, a larger fraction of the methanol consumed was converted to energy, reducing the observed growth yield.     

Exposure of the marine deposit feeder Hydrobia ulvae to sediment spiked with LAS congeners

The lethal and sub-lethal toxicity of LAS congeners to the mollusc gastropod Hydrobia ulvae were assessed in spiked sediment bioassays. This complements the little knowledge available to date on mixture effects in the sediment compartment. The LAS homologues joint effect was concentration additive (∑TU_i = 0.8-1). As opposed to the 10-d LC10 based on the sediment associated LAS concentration (91-330 mg/kg) which was independent of the homologue chain length, the LC10 based on the dissolved LAS fraction (0.804-0.068 mg/L) decreased as the homologue chain length increased from 10 to 13 carbons. The quantitative structure-activity relationship (QSAR) derived from these data was log (1/LC10 (mol/L)) = 0.64 log K_ow + 4.40 (n = 5; r² = 0.76; s = 0.24). It showed an apparent higher toxicity compared to the typical QSAR for polar narcosis in water-only systems probably due to the simultaneous exposure of the snail to LAS through the dissolved and the sediment associated fractions. The egestion rate of the surviving snails recovered after few days' exposure (1-d NOEC: 40-107 mg/kg, 9-d NOEC: 65-190 mg/kg) which suggests that the organisms were able to acclimate to LAS during the exposure.     

Toxicity of chlorine to juvenile spot, Leiostomus Xanthurus

The sensitivity of juvenile spot, Leiostomus xanthurus, to total residual chlorine (TRC) in flowing sea-water was investigated. Incipient LC₅₀ bioassays, histopathology, avoidance tests and the combined effect of thermal stress and TRC were used to assess sensitivity.

Estimated incipient LC₅₀ values were 0.12 mg 1⁻¹ TRC at 10°C and 0.06 mg 1⁻¹ TRC at 15°C. Histological examination of spot used in the incipient LC₅₀ bioassay at 15°C and sacrificed while alive indicated pseudobranch and gill damage occurred in individuals exposed to a measured TRC concentration of 1.57 mg 1⁻¹. Spot exposed to lower concentrations of TRC, 0.02 0.06 mg 1⁻¹ at 15°C and sacrificed alive showed no consistent tissue damage.

Spot demonstrated temperature dependent avoidance responses to TRC. At 10°C, a concentration of 0.18 mg 1⁻¹ was required for significant (X²; P < 0.05) avoidance; at 15 and 20°C, spot showed significant avoidance of TRC concentrations as low as 0.05 mg 1⁻¹.

Simultaneous exposure of spot to thermal stress (5, 10 or 13°C above the acclimation temperature of 15°C) at measured TRC concentrations of 0.05 0.07 and 0.34–0.52 mg 1⁻¹ demonstrated a significant, (Z²) with Yates correction, P < 0.05) increase in sensitivity to TRC with increased temperature and exposure times for some of the groups tested.     

Occurrence and behaviour of linear alkylbenzenesulphonates, nonylphenol, nonylphenol mono- and nonylphenol diethoxylates in sewage and sewage sludge treatment

The fluxes of linear alkylbenzenesulphonates (LAS), nonylphenol (NP), nonylphenol monoethoxylate (NP1EO) and nonylphenol diethoxylate (NP2EO) through sewage and sludge treatment of 29 Swiss sewage treatment plants were investigated. Reversed-phase high-performance liquid chromatography (HPLC) was used to determine LAS. Normal-phase HPLC was employed to measure NP, NP1EO and NP2EO which are metabolites of the nonionic surfactants of the nonylphenol polyethoxylate type (NPnEO). Quantitative determinations were performed of raw sewage, primary and secondary effluents and of sewage sludge. Under normal conditions of sewage and sludge treatment, LAS were efficiently removed from the raw wastewater (> 99% w/w) and were partly transferred to the sewage sludge (15–20% w/w). About 50% (molar base) of NPnEO in the sewage were transformed to NP and accumulated in the digested sludge. Large variations existed among different sewage treatment plants. It was estimated that 1.0 g m⁻² y⁻¹ of LAS and 0.3 g m⁻² y⁻¹ of NP are applied with sewage sludge to Swiss soils.     

复合荷载作用下简支钢梁弹性弯扭屈曲研究

为建立复合荷载作用下简支钢梁弹性弯扭屈曲的设计理论,提出不同类型、不同作用点的横向荷载和不等端弯矩共同作用下的弯扭屈曲总势能方程。采用Rayleigh-Ritz法推导复合荷载作用下临界弯矩M_cr的通用计算式及系数（参数）C₁′、C₂′a、C₃′的计算式,并得到复合荷载作用的C₁′、C₂′a、C₃′与单一荷载作用的系数C₁、C₂、C₃的关系。采用Rayleigh-Ritz法推导临界荷载的通用计算式,并通过数值算例验证其正确性,最后分析钢梁自重对弯扭屈曲临界荷载的影响。研究表明：复合荷载作用下的临界弯矩Mcr通用计算式可完全采用单一荷载作用下的M_cr通用计算式表示;钢梁自重对其弯扭屈曲临界荷载的影响较小,计算中可忽略不计。     

Effects of substrate concentrations on the growth of heterotrophic bacteria and algae in secondary facultative ponds

This paper presents the effect of substrate concentration on the growth of a mixed culture of algae and heterotrophic bacteria in secondary facultative ponds (SFPs) utilizing settled domestic sewage as a sole source of organic carbon. The growth of the mixed culture was studied at the concentrations ranging between 200 and 800 mg COD/l in a series of batch chemostat reactors. From the laboratory data, the specific growth rate (micro) was determined using the modified Gompertz model. The maximum specific growth rate ( micro(max)) and half saturation coefficients (K(s)) were calculated using the Monod kinetic equation. The maximum observed growth rate ( micro(max)) for heterotrophic bacteria was 3.8 day(-1) with K(s) of 200 mg COD/l. The micro(max) for algal biomass based on suspended volatile solids was 2.7 day(-1) with K(s) of 110 mg COD/l. The micro(max) of algae based on the chlorophyll-a was 3.5 day(-1) at K(s) of 50mg COD/l. The observed specific substrate removal by heterotrophic bacteria varied between the concentrations of substrate used and the average value was 0.82 (mg COD/mg biomass). The specific substrate utilization rate in the bioreactors was direct proportional to the specific growth rate. Hence, the determined Monod kinetic parameters are useful for the definition of the operation of SFPs.     

钢梁弯扭屈曲临界弯矩通用公式研究

为研究钢梁弯扭屈曲总势能方程中的各势能项对临界弯矩M_cr通用公式中各系数取值的影响,由总势能方程推导临界弯矩M_cr的通用公式及3种典型总势能方程下各系数的计算式,揭示各势能项与各个系数的联系,计算无侧向支撑简支钢梁临界弯矩M_cr通用公式中各系数的数值,并分别分析现行国家标准GB 50018-2002和GB 50017-2003的整体稳定系数φ_b设计公式存在取值各异的问题。结果表明：系数C₁、C₂和C₃的取值除都与非线性正应变能和非线性剪应变能有关外,系数C₁的取值还与圣维南扭转应变能有关,系数C₂的取值还与外荷载势能有关,而影响系数C₃取值的势能项则因总势能方程而异;现行国家标准GB 50018-2002和GB 50017-2003的φ_b设计公式应采用统一的形式,且应统一φ_b设计公式中系数C₃的取值。     

Hydrogen production using single-chamber membrane-free microbial electrolysis cells

Microbial electrohydrogenesis provides a new approach for hydrogen generation from renewable biomass. Membranes were used in all the reported microbial electrolysis cells (MECs) to separate the anode and cathode chambers. To reduce the potential losses associated with membrane and increase the energy recovery of this process, single-chamber membrane-free MECs were designed and used to investigate hydrogen production by one mixed culture and one pure culture: Shewanella oneidensis MR-1. At an applied voltage of 0.6V, this system with a mixed culture achieved a hydrogen production rate of 0.53m(3)/day/m(3) (0.11m(3)/day/m(2)) with a current density of 9.3A/m(2) at pH 7 and 0.69m(3)/day/m(3) (0.15m(3)/day/m(2)) with a current density of 14A/m(2) at pH 5.8. Stable hydrogen production from lactic acid by S. oneidensis was also observed. Methane was detected during the hydrogen production process with the mixed culture and negatively affected hydrogen production rate. However, by employing suitable approaches, such as exposure of cathodes to air, the hydrogenotrophic methanogens can be suppressed. The current density and volumetric hydrogen production rate of this system have potential to increase significantly by further reducing the electrode spacing and increasing the ratio of electrode surface area/cell volume.     

Production, composition and Pb2+ adsorption characteristics of capsular polysaccharides extracted from a cyanobacterium Gloeocapsa gelatinosa

Pb²⁺ adsorption by the living cells of the cyanobacterium Gloeocapsa gelatinosa was studied. Cyanobacterial cells with intact capsular polysaccharide (CPS) showed 5.7 times higher Pb adsorption capacity than that of cells without CPS. The adsorbed Pb was desorbed by EDTA, indicating that Pb²⁺ adsorption occurred mainly on cell surface. Production, sugar content and ability of CPS to remove Pb²⁺ were then studied in details. CPS production by G. gelatinosa increased when culture time was prolonged. The maximum CPS production was 35.43 mg g⁻¹ dry weight after 30-day cultivation. Xylose, arabinose, ribose, rhamnose, galactose, glucose, mannose and fructose were the neutral sugars presented in CPS of G. gelatinosa. Acidic sugars including galacturonic and glucuronic acids were also found in CPS. The amount and composition of G. gelatinosa's CPS varied according to its growth phase and culture conditions. The highest amount of acidic sugars was produced when cultured under low light intensity. The extracted CPS rapidly removed Pb²⁺ from the solution (82.22±4.82 mg Pb²⁺ per g CPS), directly demonstrating its roles in binding Pb²⁺ ions. Its ability to remove Pb²⁺ rapidly and efficiently, to grow under sub-optimal conditions (such as low pH and low light intensity), and to produce high amount of CPS with acidic sugars, leads us to conclude that G. gelatinosa is a potential viable bioadsorber for mildly acidic water contaminated with Pb²⁺.     

Survie in vitro de Yersinia dans les eaux d'alimentation et recuperation par filtration sur membraneIn vitro survival of Yersinia strains in drinking water and recovery by membrane filtration

This study was undertaken to determine the survival characters and optimal parameters for the recovery of Yersinia from drinking water samples. Experiments were done by artificial contamination of 4 different water samples with 4 Yersinia strains.

The strains tested along this study are: Y. enterocolitica 4052, 0:3 and 4360, 0:9, Y. enterocolitica intermedia 3953, 0:17 and Y. pseudotuberculosis 0:1. The water samples are characterized by an increasing mineralization: Distillated water, Gerardmer water (-30,300 ω cm⁻¹—Hardness = 1° Français), Nancy water (-4500 ω cm⁻¹—H = 10°) and Vittel water (-730 ω cm⁻¹, H = 71°).

Total mineralization does not significantly affect the survival of Yersinia at 20°C, but causes a dramatic decrease of the Yersinia strains at 4°C for the most mineralized water. There is no pH effect from alkaline, neutral and acidic values until pH 4.4 but Yersinia is greatly affected by more acidic pH.

The previous stay of Yersinia strains in artificially contaminated water causes a reduction of kinetic growth during cultivation on peptone-water but does not change the recovery rate by filtration method.

The recovery rate of the filtration method using cellulose acetate membranes (mean porosity = 0.45 μm) is very poor, ranging from 0 to 8%. The efficiency is significantly higher with the aid of polyester membranes treated by laser (Nucleopore−0.40 μm) ranging from 12 to 53% according to the mineralization of the water. The best recovery rate is obtained with the moderately mineralized water.

These results can explain the poor results obtained with the use of water bacteriology classical methods for the search of Yersinia.

Thus the best results can be predicted from the concentration of fairly mineralized water by filtration on polyester membrane at neutral, alkaline or weakly acidic pH.     

A comparison of fenuron degradation by hydroxyl and carbonate radicals in aqueous solution

A comparative study of the transformation of the herbicide fenuron (1,1-dimethyl-3-phenylurea) by hydroxyl radicals and carbonate radicals in aqueous solution (pH 7.2-phosphate buffer) has been undertaken. Hydroxyl radical was generated by the well-known photolysis of hydrogen peroxide at 254 nm and carbonate radical was formed by photolysis of Co(NH₃)₅CO₃⁺ at 254 nm. Competitive kinetic experiments were performed with atrazine used as the main competitor for both processes. Accordingly, the second-order rate constant of reaction between fenuron and carbonate radical was found to be (7−12±3)×10⁶ M⁻¹ s⁻¹ [(7±1)×10⁹ M⁻¹ s⁻¹ for hydroxyl radical]. The formation of degradation products was studied by LC-MS in the two cases and a comparison has been performed. The reaction with carbonate radical leads to the formation of a quinone-imine derivative which appears as the major primary product together with ortho and para hydroxylated compounds. These two compounds represent the major products in the reaction with hydroxyl radicals. The reaction of both radicals also leads to the transformation of the dimethylurea moiety.     

Development of an immunoassay for a quaternary ammonium compound, benzyldimethyldodecylammonium chloride

A competitive enzyme-linked immunosorbent assay (CELISA), based on polyclonal antibodies, was developed to measure benzyldimethyldodecylammonium chloride (BDD₁₂AC), a component of benzalkonium chloride (BAK). The polyclonal antibodies recognized free benzyldimethyldodecylammonium bromide (BDD₁₂AB) in an indirect competitive ELISA with a 50% inhibition (IC₅₀) of 0.66±0.20 μg/ml and a detection limit of 0.043±0.033 μg/ml. The two other components of BAK, benzyldimethyltetradecylammonium chloride (BDT₁₄AC) and benzyldimethylhexadecylammonium chloride (BDH₁₆AC), were recognized to varying degrees by the antibodies. The antibodies did not cross-react with other compounds such as fatty acids and alcohols, amino acids, amines, and short-chain quaternary ammonium compounds. Cross-reactivity was observed with other alkyldimethylbenzylammonium compounds (ADBACs). The CELISA and HPLC were used to measure the amount of pure BDD₁₂AB spikes in milk and BDD₁₂AC in five commercial products. HPLC analysis correlated well with CELISA analysis for spiked samples.